基于网络药理学与分子对接和实验验证探讨费菜抗炎作用的分子机制

Molecular Mechanism of Phedimus aizoon(Linnaeus)'t Hart.on Antiinflammatory Effect Based on Network Pharmacology and Molecular Docking and Experiment Research

目的:本文基于网络药理学技术与分子对接和实验验证探讨费菜抗炎作用机制。方法:利用TCMSP、HERB等数据库,以口服生物利用度≥30%为筛选条件,得到费菜药效成分及其潜在靶点。利用GeneCards、OMIM等数据库查找炎症相关靶点,通过STRING数据库和Cytoscape3.8.0软件绘制PPI蛋白互作图,分析费菜中抗炎的关键药效成分和潜在作用靶点。利用David数据库对潜在作用靶点进行GO富集和KEGG信号通路富集,并进行可视化分析。应用AutoDock软件将药效成分与核心靶点进行分子对接,并进行可视化分析。利用高效液相色谱法分析药效成分。采用LPS诱导的RAW264.7细胞炎症模型,验证药效成分的抗炎作用。结果:筛选得到5个药效成分及相关靶点130个,抗炎靶点421个,两者取交集后获得费菜潜在抗炎靶点22个,其中关键靶点有PPARG、EGFR、TP53等,发挥关键作用的主要成分为β-谷甾醇和没食子酸。GO及KEGG富集分析结果表明,β-谷甾醇和没食子酸主要通过调控聚合酶II启动子的转录正调控、神经元凋亡过程的正调控和脂多糖反应等生物过程,通过癌症通路、乙型肝炎、TNF信号通路、MAPK信号通路等信号通路发挥抗炎作用。分子对接结果显示抗炎药效成分均与靶点蛋白分子对接结果良好。高效液相色谱法证明费菜中含有药效成分β-谷甾醇和没食子酸。实验验证结果表明,β-谷甾醇和没食子酸可显著降低促炎因子NO含量,提高抗炎因子IL-10含量(P<0.05),发挥抗炎作用。结论:本研究揭示了费菜通过调控多靶点、多途径发挥抗炎作用,为费菜的研究和应用奠定了基础。

Objective:To study the molecular mechanism of Phedimus aizoon(Linnaeus)'t Hart.on anti-inflammatory effect by network pharmacology,molecular docking,and experiment research.Methods:The potential anti-inflammatory effect ingredients and targets were acquired from TCMSP,HERB,and other databases under the screening condition of oral bioavailability≥30%.The inflammation-related targets were collected from GeneCards,OMIM,and other databases,and the protein-protein interaction(PPI)analysis to obtain the key anti-inflammatory effect targets.Meanwhile,GO annotation and KEGG signal pathway enrichment analysis of the key targets was analyzed using the David database.Moreover,this study verified the key targets predicted by AutoDock molecular docking.The key anti-inflammatory effect ingredients of Phedimus aizoon(Linnaeus)'t Hart.were analyzed using high-performance liquid chromatography(HPLC).The anti-inflammatory effect of the key ingredients was verified on the LPS-induced RAW264.7 cell model.Results:The results showed that there were five potent components corresponding to 130 related targets,and 421 inflammation-related targets,with 22 intersection anti-inflammatory targets,among which the key targets were PPARG,EGFR,TP53,etc.β-sitosterol and gallic acid(3,4,5-trihydroxy benzoic acid)were the main ingredients that can be anti-inflammatory.GO annotation and KEGG signal pathway enrichment analysis indicated thatβ-sitosterol and gallic acid played an anti-inflammatory role through multiple pathways such as transcription from RNA polymerase II promoter,positive regulation of neuron apoptotic process,and response to lipopolysaccharide,etc.,as well as signal pathways including pathways in cancer,hepatitis B,TNF signaling pathway,MAPK signaling pathway,and other signaling pathways.Furthermore,the molecular docking analysis revealed that the key targets had good binding affinities with the key active ingredients.β-sitosterol and gallic acid as active ingredients in Phedimus aizoon(Linnaeus)'t Hart.were analyzed by HPLC.The experimental results showed thatβ-sitosterol and gallic acid could significantly reduce NO content,increase the content of IL-10(P<0.05),and play an anti-inflammatory effect.Conclusion:This study revealed that Sedum aizoon L.would play an anti-inflammatory role by regulating multiple targets and multiple pathways,which laid a foundation for the research and application of Phedimus aizoon(Linnaeus)'t,Hart.